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Anti-PODXL 抗体 [EPR9518] - BSA and Azide free (ab269888)

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Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Flow Cytometry - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunocytochemistry/ Immunofluorescence - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
  • Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9518] to PODXL - BSA and Azide free
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human

製品の概要

  • 製品名

    Anti-PODXL antibody [EPR9518] - BSA and Azide free
    PODXL 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR9518] to PODXL - BSA and Azide free

  • 由来種

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • 免疫原

    Recombinant fragment within Human PODXL aa 300-500. The exact sequence is proprietary.
    Database link: O00592

  • ポジティブ･コントロール

    • WB: HCT116, HAP1, Raji and HeLa cell lysates. Human fetal kidney tissue lysate. IHC-P: Human kidney tissue. Human hepatocellular carcinoma, breast carcinoma and endometrial carcinoma tissue. Human glioma. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.

  • 特記事項

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    ab269888 is the carrier-free version of ab150358. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab269888 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

製品の特性

アプリケーション

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab269888 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

アプリケーション Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
WB
Human
アプリケーション Abreviews 特記事項
WB
Use at an assay dependent concentration. Detects a band of approximately 165 kDa (predicted molecular weight: 58 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.

 
特記事項
WB
Use at an assay dependent concentration. Detects a band of approximately 165 kDa (predicted molecular weight: 58 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.

 
  • 追加情報
    Is unsuitable for IP.

    • ターゲット情報

    • 機能

      Involved in the regulation of both adhesion and cell morphology and cancer progression. Function as an anti-adhesive molecule that maintains an open filtration pathway between neighboring foot processes in the podocyte by charge repulsion. Acts as a pro-adhesive molecule, enhancing the adherence of cells to immobilized ligands, increasing the rate of migration and cell-cell contacts in an integrin-dependent manner. Induces the formation of apical actin-dependent microvilli. Involved in the formation of a preapical plasma membrane subdomain to set up inital epithelial polarization and the apical lumen formation during renal tubulogenesis. Plays a role in cancer development and aggressiveness by inducing cell migration and invasion through its interaction with the actin-binding protein EZR. Affects EZR-dependent signaling events, leading to increased activities of the MAPK and PI3K pathways in cancer cells.

    • 組織特異性

      Glomerular epithelium cell (podocyte).

    • 配列類似性

      Belongs to the podocalyxin family.

    • ドメイン

      Both the O-glycan-rich domain of the extracellular domain and the C-terminus PDZ-binding motif (DTHL) in the cytoplasmic tail harbor an apical sorting signal. The cytoplasmic domain is necessary for the apical membrane targeting and renal tubulogenesis. The cytoplasmic C-terminus PDZ-binding motif (DTHL) is essential for interaction with SLC9A3R1 and for targeting SLC9A3R1 to the apical cell membrane. The extracellular domain is necessary for microvillus formation (By similarity). The large highly anionic extracellular domain allows to maintain open filtration pathways between neighboring podocyte foot processes.

    • 翻訳後修飾

      N- and O-linked glycosylated. Sialoglycoprotein.

    • 細胞内局在

      Apical cell membrane. Cell projection, lamellipodium. Cell projection, filopodium. Cell projection, ruffle. Cell projection, microvillus. Membrane raft. Membrane. In single attached epithelial cells is restricted to a preapical pole on the free plasma membrane whereas other apical and basolateral proteins are not yet polarized. Colocalizes with SLC9A3R2 at the apical plasma membrane during epithelial polarization. Colocalizes with SLC9A3R1 at the trans-Golgi network (transiently) and at the apical plasma membrane. Its association with the membrane raft is transient. Colocalizes with actin filaments, EZR and SLC9A3R1 in a punctate pattern at the apical cell surface where microvilli form. Colocalizes with EZR and SLC9A3R2 at the apical cell membrane of glomerular epithelium cells (By similarity). Forms granular, punctuated pattern, forming patches, preferentially adopting a polar distribution, located on the migrating poles of the cell or forming clusters along the terminal ends of filipodia establishing contact with the endothelial cells. Colocalizes with the submembrane actin of lamellipodia, particularly associated with ruffles. Colocalizes with vinculin at protrusions of cells. Colocalizes with ITGB1. Colocalizes with PARD3, PRKCI, EXOC5, OCLN, RAB11A and RAB8A in apical membrane initiation sites (AMIS) during the generation of apical surface and luminogenesis (By similarity).
    • Information by UniProt

    • 参照データベース

      • 製品の状態

        There are 2 isoforms produced by alternative splicing.

      • 別名

        • GCTM-2 antigen antibody
        • Gp2 antibody
        • Gp200 antibody
        • MGC138240 antibody
        • PC antibody
        • PCLP antibody
        • PCLP-1 antibody
        • PCLP1 antibody
        • Pcx antibody
        • Podocalyxin antibody
        • Podocalyxin like antibody
        • Podocalyxin like protein antibody
        • Podocalyxin-like protein 1 antibody
        • Podxl antibody
        • PODXL_HUMAN antibody
        see all

      画像

      • Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        All lanes : Anti-PODXL antibody [EPR9518] (ab150358) at 1/10000 dilution

        Lane 1 : Wild-type HeLa cell lysate
        Lane 2 : PODXL knockout HeLa cell lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 58 kDa
        Observed band size: 160 kDa
        why is the actual band size different from the predicted?



        This data was developed using the same antibody clone in a different buffer formulation (ab150358).

          Lanes 1- 2: Merged signal (red and green). Green - ab150358 observed at 160 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

         ab150358 was shown to react with PODXL in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264984 (knockout cell lysate ab257210) was used. Wild-type HeLa and PODXL knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab150358 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunohistochemical analysis of paraffin embedded human breast adenocarcinoma tissue using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        All lanes : Anti-PODXL antibody [EPR9518] (ab150358) at 1/1000 dilution

        Lane 1 : Wild-type HCT116 cell lysate
        Lane 2 : PODXL knockout HCT116 cell lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 58 kDa
        Observed band size: 200 kDa why is the actual band size different from the predicted?



        This data was developed using the same antibody clone in a different buffer formulation (ab150358).

        Lanes 1- 2: Merged signal (red and green). Green - ab150358 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

         ab150358 was shown to react with PODXL in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266887 (knockout cell lysate ab257211) was used. Wild-type HCT116 and PODXL knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab150358 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

      • Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        All lanes : Anti-PODXL antibody [EPR9518] (ab150358) at 1/1000 dilution

        Lane 1 : Wild-type HAP1 whole cell lysate
        Lane 2 : PODXL knockout HAP1 whole cell lysate
        Lane 3 : Raji whole cell lysate
        Lane 4 : HeLa whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Predicted band size: 58 kDa



        Lanes 1 - 4: Merged signal (red and green). Green - ab150358 observed at 160 kDa. Red - loading control, ab9484, observed at 37 kDa.

        ab150358 was shown to specifically react with PODXL in wild-type cells as signal was lost in PODXL knockout cells. Wild-type and PODXL knockout samples were subjected to SDS-PAGE. Ab150358 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Flow Cytometry - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Flow Cytometry - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling PODXL with purified ab150358 at 1/250 dilution (red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Immunocytochemistry/ Immunofluorescence - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunocytochemistry/ Immunofluorescence - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labeling PODXL with purified ab150358 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei couterstained with DAPI (blue).

        Secondary Only Control: PBS was used instead of the primary antibody as the negative control. 

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunohistochemical analysis of paraffin embedded human glioma tissue using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunohistochemical analysis of paraffin embedded normal human kidney tissue using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunohistochemical analysis of paraffin embedded human skeletal muscle tissue using unpurified ab150358 showing negative staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunohistochemical analysis of paraffin embedded human endometrial carcinoma tissue using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunohistochemical analysis of paraffin embedded human hepatocellular carcinoma vessels using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunohistochemical analysis of paraffin embedded human kidney tissue labeling PODXL with unpurified ab150358 antibody at a dilution of 1/100. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling PODXL with purified ab150358 at 1/1000 dilution (0.44 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) is the secondary antibody.

        PBS instead of the primary antibody was used as the negative control.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

      • Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
        Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

      プロトコール

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      参考文献 (0)

      ab269888 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      ab269888 は論文での使用が確認できていません。

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